Screening and improvement of an anti-VEGF DNA aptamer.

Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry
Yoshihiko NonakaKazunori Ikebukuro

Abstract

To obtain an aptamer with a high affinity for vascular endothelial growth factor (VEGF), we focused on the receptor-binding domain (RBD) of VEGF as a target epitope. Three rounds of screening gave Vap7, which bound to the VEGF isoforms VEGF(121) and VEGF(165) with K(D) values of 1.0 nM and 20 nM, respectively. Moreover, Vap7 showed specificity within the VEGF family. Secondary structure predictions and circular dicrhoism suggested that Vap7 folds into a G-quadruplex structure. We obtained a mutant aptamer that contains only this region of the aptamer sequence. This truncated mutant (V7t1) bound to both VEGF(121) and VEGF(165) with K(D) values of 1.1 nM and 1.4 nM, respectively. Its sequence was 5'-TGTGGGGGTGGACGGGCCGGGTAGA-3', and it appeared to form a G-quadruplex structure. We also produced an aptamer heterodimer consisting of our previously derived aptamer (del5-1), which binds to the heparin-binding domain of VEGF, linked to V7t1. The resulting heterodimer bound strongly to VEGF(165) with a K(D) value of 4.7 x 10(2) pM.

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Citations

Apr 10, 2013·Pharmaceutical Research·Nishant Kumar JainIpsita Roy
Jun 16, 2012·Analytical Chemistry·Kaori TsukakoshiKazunori Ikebukuro
Jul 4, 2012·Analytical Chemistry·Ronit FreemanItamar Willner
Dec 15, 2012·Analytical Chemistry·Yoshihiko NonakaKazunori Ikebukuro
Jun 12, 2013·PloS One·Wataru YoshidaKazunori Ikebukuro
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Jun 22, 2017·Analytical and Bioanalytical Chemistry·Kyung Hyun Lee, Huaqiang Zeng
Aug 26, 2017·International Journal of Molecular Sciences·Xiaoqiu WuGe Zhang
Apr 5, 2018·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Minhee KimDong-Eun Kim
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Mar 18, 2020·Chembiochem : a European Journal of Chemical Biology·Sepehr ManochehryYingfu Li

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Methods Mentioned

BETA
biosensors
SELEX
surface plasmon resonance
chip
circular dichroism
PCR
Cytomation
chips
biosensing

Software Mentioned

QGRS Mapper
Biacore T100
fold

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